Investigation into genetic variability of Hazel Grouse Bonasa bonasia (=Tetrastes bonasia) population in Lithuania using non-invasive sampling

摘要

We analysed 51 sequences of the Hazel Grouse's (Bonasa bonasia) mitochondrial control region (CR) to obtain data on the variability of the fragment of the selected size and to assess its suitability for the evaluation of the genetic structure of the species population in Lithuania. The samples (feathers and faeces) used for DNA extraction were collected non-invasively from different localities of Lithuania in 2008-2010. In order to amplify the short but variable region of the left domain in the CR ofmitochondrial DNA, a new primer pair (BON-F and BON-R) specific to the Hazel Grouse was designed. The selected 116 bp long fragment of the mitochondrial CR exhibited very high polymorphism. Among 51 novel sequences from Lithuania, 23 variable sites and21 haplotypes with frequency values ranging from 1.96 to 17.65% were defined. The haplotype diversity (H) and nucleotide diversity (k) among haplotypes were 0.928 (±0.018) and 0.03955 (±0.00277), respectively. In order to compare genetic variability among Hazel Grouse populations, the newly obtained CR sequences, representing the Lithuanian population, were combined with sequences derived from GenBank, including 21 haplotypes from Poland. The haplotype network analysis revealed a complicated evolutionary history of the species. The neighbour-joining tree derived from 42 Hazel Grouse haplotypes shows a clear separation of two clades of evolutionarily distinct haplogroups both in Lithuania and Poland and some evidence of clustering related to samplinglocations.